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. 2019 Apr 10;9:5858. doi: 10.1038/s41598-019-42415-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Double immunofluorescence staining of human tongue tissue sections. (A–C) CD34 (green) and CD31 (red) immunostaining with 4′,6-diamidino-2-phenylindole (DAPI; blue) counterstain for nuclei. Telocytes (TCs)/CD34+ stromal cells in the tongue lamina propria (A,B) and skeletal muscle (C) lack CD31 immunoreactivity. The endothelial cells of blood vessels (arrows) are CD34+/CD31+ (A,C). At variance with double stained blood vessels (BV), the endothelium of lymphatic vessels (LV) is CD34−/CD31+ (B). Some leukocytes are also CD31+ (A,B). (D–F) CD34 (green) and α-smooth muscle actin (α-SMA; red) immunofluorescence labeling with DAPI counterstain. CD34+ TCs do not coexpress α-SMA. (D) CD34+ TCs envelop secretory salivary gland units outside of α-SMA + myoepithelial cells. (E,F) CD34+ TCs externally encircle the vascular smooth muscle cell layer of arterioles (arrows; higher magnification in the inset). (G–I) CD34 (green) and c-kit/CD117 (red) immunofluorescence with DAPI counterstain. The CD34+ TC meshwork is immunophenotypically negative for the c-kit/CD117 marker. c-kit/CD117 is detectable only in oval/round-shaped mast cells (arrows) often in close relationship with CD34+ TC processes (higher magnification in the inset). Scale bar: 50 µm (A–I).